CN102747017A - Bacillus amyloliquefaciens and application thereof - Google Patents
Bacillus amyloliquefaciens and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of agriculture microbe and discloses bacillus amyloliquefaciens and application of the bacillus amyloliquefaciens. The rhizosphere growth promoting rhinoacteria JX1 is named as bacillus amyloliquefaciens (bacillus amyloliquefaciens) in class, and conserved in the common microbe center of China microbial culture collection management committee on Dec 20, 2011, and the conservation number is CGMCC No.5624. The rhizosphere growth-promoting rhinoacteria JX1 (CGMCC No.5624) can produce heteroauxin with high yield and grow by utilizing insoluble metasilicate containing potassium as a potassium source. Therefore, the bacillus amyloliquefaciens JX1 can be applied to promoting plant growth.
Description
Technical field
The invention belongs to agriculture microorganism field, relate to a kind of bacillus amyloliquefaciens and application.
Background technology
Moisture soil is that fluvial deposit is influenced by ground water movement and farming activity and the soil that forms, gains the name because of the Evening Tide phenomenon is arranged.In China, be distributed in the Yellow River more, the river plain in downstream and on the south in the plains region and the Yangtze valley in Jiangsu, Anhui, river, lake plain and the area, delta in downstream.Moisture soil range of distribution physical features is smooth, and soil layer is deep, and hydrothermal resources is abundanter, and it is wide to make kind of property, is the main upland soil of China, abounds with grain and cotton.But the Yellow River and Huai He River sea plain that the moisture soil distribution area is maximum, drought and waterlogging happens occasionally, and saline and alkaline harm is still arranged, and soil nutrient is low or lack in addition, and in most of the genus, low productive soil, crop yield is low and unstable.Must strengthen the reasonable utilization and the improvement of moisture soil.
Rhizosphere is urged living bacterium (Plant Growth Promoting Bacteria, be called for short PGPB) and is defined as the free living that the helps plant-growth under certain condition bacterium at soil, rhizosphere, root table, phyllosphere.These bacteriums can fixed nitrogen, dissolve phosphorus, dissolve iron, and produce plant hormone, like growth hormone, Plant hormones regulators,gibberellins, phytokinin and ethene.In addition, they can also improve the resistance of plant, comprise arid, high salt, heavy metallic poison and agricultural chemicals.Therefore from moisture soil, separate obtaining short living bacterium of rhizosphere and crop formation syntaxial system, utilize biological prosthetic to improve moisture soil, become the focus of current research.
Summary of the invention
An object of the present invention is to provide a kind of bacillus amyloliquefaciens.
Another object of the present invention provides the application of this bacillus amyloliquefaciens.
The object of the invention can be realized through following technical scheme:
Rhizosphere is urged living bacterium JX1, classification called after bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and on December 20th, 2011 was preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, and preserving number is CGMCC No.5624.
Short living bacterium JX1 (CGMCC No.5624) bacterium colony of rhizosphere is less to be white, protuberance, neat in edge, and smooth surface is moistening, and is opaque, and gemma is produced in irregular shaft-like arrangement.
The physio-biochemical characteristics of the short living bacterium JX1 (CGMCC No.5624) of rhizosphere are: Gram-positive, and amphimicrobian, chemoheterotrophy, catalase is positive; M.R tests positive, and VP tests negative, and the starch hydrolysis is positive; Gelatin liquefaction positive, nitrate reduction is positive, and Citrate trianion utilizes positive.
The major nitrogen source that the short living bacterium JX1 (CGMCC No.5624) of rhizosphere uses when cultivating includes but not limited to peptone, yeast powder, L-Ala, saltpetre, an ammonium nitrate, ammonium sulfate, urea; The main carbon source of using includes but not limited to glucose, sucrose, fructose, wood sugar, N.F,USP MANNITOL, lactose, SANMALT-S; The inorganic component that uses includes but not limited to Repone K, sodium-chlor, SODIUM PHOSPHATE, MONOBASIC, potassium hydrogenphosphate, tricalcium phosphate, Calcium dichloride dihydrate, bitter salt, seven water and ferrous sulfate.Bacillus amyloliquefaciens JX1 (CGMCC No.5624) fermentation can be carried out under the environment of pH5 ~ 9 at 28 ~ 32 ℃.
Described preserving number is the application of bacillus amyloliquefaciens JX1 in promoting plant-growth of CGMCC No.5624.
Described plant optimization peanut.
Said rhizosphere short living bacterium JX1 (CGMCC No.5624) ability high yield indolylacetic acid also can utilize insoluble to contain potassium silicate and grow for the potassium source.
Described preserving number is the application of bacillus amyloliquefaciens JX1 in peanut cultivation of CGMCC No.5624.
The ability of short living bacterium JX1 (CGMCC No.5624) the secretion indolylacetic acid of rhizosphere of the present invention (IAA) is strong, reaches 30.60 μ gmL
-1Indolylacetic acid is a kind of of plant hormone, can promote the growth of root.Produce the bacterial classification of indolylacetic acid,, produce physiological process and metamorphosis that plant hormones such as IAA and a small amount of GA3 influence plant when utilizing plant metabolism to produce secretory product often attached to root system of plant or leaf surface.Show as the elongation of direct promotion root, thus increased with soil in the chance that contacts of nutritive substance; Can improve the content of plant materials Endogenous IAA; Inducing plant defence expression of gene, it is disease-resistant to improve plant materials, resistance such as drought resisting.As prioritization scheme of the present invention, the fermentation of short the livings bacterium JX1 of said rhizosphere time is carried out in pH8 ~ 9, and this environment produces IAA down and measures the highest.
As further optimization of the present invention, the carbon source that the short living bacterium JX1 (CGMCC No.5624) of said rhizosphere adopts is a lactose, and the nitrogenous source of employing is yeast powder or peptone or both combinations.The substratum that utilizes above-mentioned carbon source and nitrogenous source to make, the amount that the short living bacterium of the rhizosphere of cultivating produces IAA is the highest.
It is to grow in the potassium source that the short living bacterium JX1 (CGMCC No.5624) of rhizosphere of the present invention contains potassium silicate with insoluble, and is translated into soluble potassium salt.Shake under bottle condition in the laboratory, the short living bacterium JX1 (CGMCC No.5624) of said rhizosphere reaches 14.23mgL to the inversion quantity of feldspar in powder
-1Explain that the JX1 bacterium has solvency action to feldspar in powder, containing potassium silicate with insoluble is to grow in the potassium source, and is translated into soluble potassium salt.
Bacterial strain JX1 point is connected to the nitrogen-free agar flat board, can grows, it possibly have certain spontaneous nitrogen fixing capacity.
Beneficial effect: rhizosphere provided by the invention is urged living bacterium JX1 (CGMCC No.5624) high yield indolylacetic acid; Can effectively insoluble be contained potassium silicate and be converted into soluble potassium salt; Improve fertilizer utilization ratio, promote plant root system development and, increase the effective potassium content of soil the absorption of fertilizer; The present invention is directed to peanut and have good promotes growth effect, the indolylacetic acid of high yield promotes growing of peanut, and the raising of the effective potassium content of soil also makes peanut higher to the utilization ratio of potash fertilizer.
Description of drawings
Fig. 1 is the bacterium colony figure of bacterial strain JX1 of the present invention;
Fig. 2 representes that different liquid amounts produce the influence of IAA to bacterial strain JX1;
Fig. 3 representes that different initial pH produce the influence of IAA to the JX1 bacterial strain;
Fig. 4 representes the influence of different carbon sources to bacterial strain JX1 product IAA;
Fig. 5 representes the influence of different nitrogen sources to JX1 bacterial strain product IAA;
Fig. 6 representes that bacterial strain JX1 contains the situation of utilizing of potassium silicate to insoluble;
Fig. 7 representes to plant peanut and inoculates the influence of JX1 bacterial strain to peanut overground part fresh weight after 30 days;
Fig. 8 representes to plant peanut and inoculates the influence of JX1 bacterial strain to the peanut plant plant height after 30 days;
Fig. 9 representes to plant peanut and inoculates the influence of JX1 bacterial strain to the peanut plant total nitrogen content after 30 days;
Figure 10 representes to plant peanut and inoculates the influence of JX1 bacterial strain to the full potassium content of peanut plant after 30 days;
Figure 11 representes to plant peanut and inoculates the influence of JX1 bacterial strain to peanut root total length after 30 days;
Figure 12 representes to plant peanut and inoculates the influence of JX1 bacterial strain to the peanut root surface area after 30 days;
Figure 13 representes to plant peanut and inoculates the influence of JX1 bacterial strain to the peanut average root diameter after 30 days;
Figure 14 representes to plant peanut and inoculates the influence of JX1 bacterial strain to peanut tip of a root number after 30 days;
Figure 15 representes to plant peanut and inoculates the influence of JX1 bacterial strain to soil IAA content after 30 days;
Figure 16 representes to plant the soil mineral nitrogen content of peanut after 30 days;
Figure 17 representes to plant the soil quick-acting potassium content of peanut after 30 days.
Biomaterial preservation information
Rhizosphere is urged living bacterium JX1; Classification called after bacillus amyloliquefaciens (Bacillus amyloliquefaciens); Be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on December 20th, 2011; The address is No. 3 Institute of Microorganism, Academia Sinica in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and preserving number is CGMCC No.5624.
Embodiment
Below in conjunction with accompanying drawing the present invention is done explanation further.
At first prepare following three kinds of substratum.
LB substratum: peptone 10g, yeast extract 5g, sodium-chlor 10g, agar 20g, zero(ppm) water 1000ml, pH7.0-7.2,121 ℃ of sterilizations, 20min.
The LB liquid nutrient medium: do not add agar, other condition is the same.
Liquid potassium bacterium substratum: sucrose 10.0g, yeast extract paste 0.5g, (NH
4)
2SO
41.0g, Na
2HPO
42.0g, MgSO
47H
2O0.5g, CaCO
31.0g, feldspar in powder 1.0g, zero(ppm) water 1000mL, 121 ℃ of sterilizations, 20min.
Nitrogen fixing capacity adopts the Ashby nitrogen-free agar: N.F,USP MANNITOL 10g, KH
2PO
40.2g, MgSO
40.2g, NaCl 0.2g, K
2SO
40.3g, CaCO
35g, zero(ppm) water 1000mL, Agar 15g, 121 ℃ of sterilizations, 20min.
Minimal medium: ammonium sulfate 2.0g; SODIUM PHOSPHATE, MONOBASIC 0.5g; Potassium hydrogenphosphate 0.5g; MAGNESIUM SULPHATE HEPTAHYDRATE 99.5 0.2g; Calcium dichloride dihydrate 0.1g, zero(ppm) water 1000mL, pH7.0,121 ℃ of sterilizations, 20min.
Moisture soil that will the slab bridge town takes from Nanjing takes by weighing the triangular flask that 10g places the 250ml that fills the 100ml aqua sterilisa, in shaking table, and 30 ℃, 150rmin
-1Vibration 20min leaves standstill 10min, obtains soil bacteria suspension-s.Contain the short living bacterium of some kinds of rhizospheres in this soil bacteria suspension-s, be applied to the LB substratum after the dilution of employing dilution method, flat board is inverted; In 30 ℃, cultivate 24h in the thermostat container after, the single bacterium colony of the dissimilar typical cases of picking; Behind dull and stereotyped purifying, 4 ℃ to be kept at the LB inclined-plane for use.
Filter out the plant growth-promoting bacterium that can secrete indolylacetic acid through qualitative test and quantitatively determined more below.
Qualitative test: with the microbionation after the separation and purification in the LB liquid nutrient medium that contains L-tryptophane (100mg/L), 30 ℃, 180rmin
-1Shaking table is cultivated 1d.Get 50 μ L bacteria suspensions and drip on the whiteware plate, add 50 μ L Salkowski color solution (50mL35%HClO simultaneously
4+ 1mL0.5M FeCl
3).With the color solution that adds 50 μ L50mg/L indolylacetic acids as positive control.The whiteware plate is observed after the room temperature lucifuge is placed 30min, and the color person of reddening representes to secrete indolylacetic acid.
Quantitatively determined: the bacterium of the secretion IAA that primary dcreening operation is obtained carries out quantitatively determined, and culture condition is the same.At first with the OD600 value of spectrophotometry bacteria suspension, then with bacteria suspension with 10000rmin
-1Centrifugal 10min gets supernatant and adds equal-volume Salkowski color solution, and lucifuge leaves standstill 30min, measures its OD
530Value.Calculate bacteria concentration OD
600Value is 1 o'clock, the content of indolylacetic acid in the unit volume fermented liquid.Analytically pure indolylacetic acid gradient dilution preparation is adopted in the drafting of typical curve.
Carrying out the screening assay of potassium decomposing situation, strains tested is inoculated in the 250mL triangular flask that fills the liquid potassium bacterium substratum of 50mL, 30 ℃, 200rmin through the product IAA bacterium qualitative, that quantitatively determined obtains
-1After cultivating 72h, get the nutrient solution 20mL that cultivates 72h, the centrifugal 20min of 6000r/min gets supernatant and measures wherein K with flame spectrophotometer
+Content.
Filter out a plant height through above mensuration and produce indolylacetic acid, the bacterial strain that ability of dissolving potassium is strong, called after JX1.As shown in Figure 1, the bacterium colony that this bacterial strain forms is less to be white, protuberance, neat in edge, and smooth surface is moistening, and is opaque, and gemma is produced in irregular shaft-like arrangement.As shown in Figure 6, it is to grow in the potassium source that bacterial strain JX1 contains potassium silicate with insoluble, and is translated into soluble potassium salt.Shake under bottle condition in the laboratory, the short living bacterium JX1 of said rhizosphere reaches 14.23mgL to the inversion quantity of feldspar in powder
-1Explain that the JX1 bacterium has solvency action to feldspar in powder, containing potassium silicate with insoluble is to grow in the potassium source, and is translated into soluble potassium salt.
The bacterial strain that the aforesaid method screening and separating is gone out; The handsome biotechnology ltd order-checking through Shanghai; Sequencing result according to 16SrDNA; Analyze in http://www.ncbi.nlm.nih.gov online query, utilize Blast software in GenBank, to carry out homology relatively, select the 16SrDNA systematic evolution tree of the sequence of close sequence and JX1 with MEGA version3 software building JX1 with other 16S rDNA sequence.According to the physiological and biochemical property of this bacterial strain, be accredited as bacillus amyloliquefaciens (Bacillus amyloliquefaciens), homology is 85%.With this bacterial strain on December 20th, 2011 in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, preserving number CGMCC No.5624.
This bacterial classification is Gram-positive, sporiferous irregular shaft-like.Bacterium colony is less to be white, protuberance, neat in edge, and smooth surface is moistening.Amphimicrobian, chemoheterotrophy.Optimum growth temperature is 30 ℃.Catalase is positive, and nitrate reduction is positive.Secretion IAA ability is strong, reaches 30.60 μ gmL
-1, containing potassium silicate with insoluble is to grow in the potassium source, and is translated into soluble potassium salt, has nitrogen fixing capacity.
The aerobic test
Pour the LB substratum of the bacterium of going out in 3 sterilized test tubes into, greatly about 2/3 place, on the aseptic technique platform, with the bacterial strain JX1 (CGMCC No.5624) of inoculating needle picking slant culture, percutaneous puncture-inoculation (must be punctured to the pipe end) in above-mentioned substratum.30 ℃ of cultivations are respectively 3 days to 7 days observationss.On the agar column surface, giving birth to the elder is aerobic bacteria, is anerobes or facultative anaerobe as giving birth to the elder along the puncture line.Test-results performance, bacterial strain JX1 (CGMCC No.5624) bacterium colony also have colony growth along the agar column surface growth in the puncture line, are amphimicrobian.
Catalatic mensuration
On clean slide, drip 1 3%H
2O
2, get bacterial strain JX1 (CGMCC No.5624) LB slant culture 1 ring that 18 ~ 24h cultivates, at H
2O
2In smear, if there have bubble to produce to be then positive, otherwise negative.Test-results shows that bacterial strain JX1 (CGMCC No.5624) is that catalase is positive.
Methyl red test (M.R test)
A. substratum and reagent peptone 5g, glucose 5g, sodium-chlor 5g, zero(ppm) water 1000mL regulates pH7.0 ~ 7.2, the packing test tube, every pipe is adorned 4 ~ 5mL, 121 ℃ of sterilization 20min.Reagent: methyl red 0.1g, 95% alcohol 300mL, zero(ppm) water 200mL.
B. spawn culture and result observe inoculating strain JX1 (CGMCC No.5624) in above-mentioned nutrient solution, cultivate 1 ~ 2 day for 30 ℃.In nutrient solution, add several methyl red reagent, present redness, be the methyl red positive, yellow negative (methyl red transformation region 4.4 redness ~ 6.0 yellow) like nutrient solution.
Test-results shows that bacterial strain JX1 (CGMCC No.5624) is that M.R is positive.
Second phthalein carbinol methine test (VP test)
A. the same methyl red test of culture medium culturing base.
B. spawn culture and result observe inoculation and cultivate same methyl red test.When doing the VP test, get nutrient solution (about 2mL) and mix mutually, add a small amount of creatine, behind the 5min that fully vibrates, redness occurs, be the VP positive like nutrient solution with the 40%NaOH of equivalent.
Test-results shows that bacterial strain JX1 (CGMCC No.5624) is that VP is negative.
The starch hydrolysis experiment
A. substratum and reagent add 0.2% Zulkovsky starch in meat soup peptone agar, the packing triangular flask, and 121 ℃ of sterilization 20min are subsequent use.Road Ge Shi iodine liquid: crystalline flake of iodine 1g, potassiumiodide 2g, earlier with a small amount of (3 ~ 5mL) dissolved in distilled water potassiumiodides add the crystalline flake of iodine at present, treat that iodine dissolves fully after, thin up is to 300mL.
B. spawn culture and result observe and to get JX1 bacterial classification (CGMCC No.5624) point and be connected on the flat board; Cultivated 3 days for 30 ℃, behind the formation bacterium colony, on flat board, drip road Ge Shi iodine liquid; To be paved with periphery of bacterial colonies degree of being; It is blue that flat board is, and periphery of bacterial colonies is irised out at present if any water white transparency, explains that starch is hydrolyzed.The size of the big or small general remark hydrolyzed starch ability of transparent circle.
Test-results shows that bacterial strain JX1 (CGMCC No.5624) is that the starch hydrolysis is positive.
The gelatin hydrolysis test
A. substratum and reagent peptone 5g, gelatin 120g, zero(ppm) water 1000mL.Regulate pH7.2 ~ 7.4, the packing test tube, the substratum height is about 4 ~ 5cm, 121 ℃ of sterilization 20min.
B. spawn culture and result observe with puncture method inoculating strain JX1 (CGMCC No.5624) in test tube central authorities.In 30 ℃ of incubators, cultivated one month, observe gelatin and whether liquefy.
Test-results shows that bacterial strain JX1 (CGMCC No.5624) is a gelatin liquefaction positive.
Nitrate reduction test
A. substratum and reagent nitrate salt liquid nutrient medium: peptone 10g, KNO
31g, zero(ppm) water 1000mL, pH7.0 ~ 7.4.Ge Lisishi (G ries) reagent: A liquid: Sulphanilic Acid 0.5g, dilute acetic acid (about 10%) 150mL; B liquid: Cai's amine 0.1g, zero(ppm) water 20mL, dilute acetic acid (about 10%) 150mL.Pentanoic reagent: pentanoic 0.5g is dissolved in the 100mL vitriol oil, uses the 20mL distilled water diluting.
B. spawn culture and result observe bacterial strain JX1 (CGMCC No.5624) are inoculated in the nitrate salt liquid nutrient medium, cultivate 1,3,5 day for 30 ℃.In white porcelain dish aperture, pour a little nutrient solution into, drip 1 reagent A and B liquid then therein respectively, when nutrient solution become pink, rose, when orange or brown etc., expression has nitrite to exist, and is that nitrate reduction is positive, otherwise negative.
The test-results bacterial strain shows that JX1 (CGMCC No.5624) is that nitrate reduction is positive.
The utilization of Citrate trianion
A. substratum and reagent Trisodium Citrate 2g, NaCl 5g, MgSO
47H
2O 0.2g, (NH
4)
2 HPO
41g, 1% Australia thymol blue aqueous solution 10mL, agar 20g, zero(ppm) water 1000mL, pH6.8-7.0,121 ℃ of sterilization 20min.
B. spawn culture and result observe and to get the JX1 bacterial classification of cultivating about 12h (CGMCC No.5624) and be inoculated on the inclined-plane, cultivate 3-7 days for 30 ℃, and substratum is the positive reaction of alkalescence (blueness) person, and constant person is then negative.
The test-results that Citrate trianion utilizes shows that bacterial strain JX1 (CGMCC No.5624) is positive.
Embodiment 3
In order further to verify the ability and the optimum condition of short living bacterium JX1 (CGMCC No.5624) product of the rhizosphere indolylacetic acid that embodiment 1 obtains, explore influence to different pH, liquid amount, different carbon source, different nitrogen sources below to indolylacetic acid output.
To contain L-tryptophane (100mg/L) LB liquid nutrient medium and press 25ml, 50ml, 75ml; 100ml, 150ml are loaded in the triangular flask of 250mL, be in the JX1 (CGMCC No.5624) of logarithmic phase by 1% (v/v) inoculum size inoculation after; Place 30 ℃, 180r.min
-1Shaking table is cultivated 24h, measures the amount of producing IAA by the method for quantitatively determined.The result is as shown in Figure 2, because bacterial strain JX1 (CGMCC No.5624) is the amphimicrobian metabolism, air flow influences the efficient that bacterial strain produces IAA, and during the 150mL liquid amount, bacterial strain produces the IAA amount at most, and along with liquid amount reduces, output is few more afterwards.
The LB substratum that will contain L-tryptophane (100mg/L) is adjusted to different pH (4,5,6,7,8,9,10) respectively; Get in the triangular flask that 150mL is loaded on 250mL; After being in the JX1 (CGMCC No.5624) of logarithmic phase by 1% (v/v) inoculum size inoculation; Place 30 ℃, 180rmin
-1Shaking table is cultivated 24h, measures the amount of producing IAA by the method for quantitatively determined, and the result is as shown in Figure 3; Show that pH is 4 and did not produce IAA at 10 o'clock, in the strong acid and strong base environment, thalline can't carry out growth metabolism; Bacterial classification produces IAA more than sour environment in little alkali environment, the ph optimum of this bacterial classification high yield IAA is 7 ~ 9.
The carbon source that in containing L-tryptophane (100mg/L) minimal medium, adds 1% (w/v) respectively; Carbon source has glucose, wood sugar, sucrose, fructose, N.F,USP MANNITOL, lactose, SANMALT-S; Get in the triangular flask that 150ml is loaded on 250ml; After being in the JX1 (CGMCC No.5624) of logarithmic phase by 1% (v/v) inoculum size inoculation, place 30 ℃, 180rmin
-1Shaking table is cultivated 24h, measures the amount of producing IAA by the method for quantitatively determined.The result is as shown in Figure 4, and this bacterial strain is when supplying with lactose, and the ability of producing IAA is the strongest, secondly is fructose, and the utilization ratio of glucose is minimum, produces IAA hardly.
The nitrogenous source that in containing L-tryptophane (100mg/L) minimal medium (not comprising ammonium sulfate), adds 0.1% (w/v) respectively; Nitrogenous source comprises an ammonium nitrate, ammonium sulfate, saltpetre, peptone, yeast powder, L-Ala, urea etc.; Get in the triangular flask that 150ml is loaded on 250ml be in the JX1 (CGMCC No.5624) of logarithmic phase by the inoculation of 1% (v/v) inoculum size after; Place 30 ℃, 180rmin
-1Shaking table is cultivated 24h, measures the amount of producing IAA by the method for quantitatively determined.The result is as shown in Figure 5, and when explaining that getting peptone is nitrogenous source, the amount of producing IAA is maximum, secondly is yeast powder, does not utilize urea.
Bacterial strain JX1 of the present invention (CGMCC No.5624) has obvious growth promoting function to peanut, describes through pot experiment below.
Gather the fresh soil of moisture soil 0 ~ 20cm soil layer under the natural condition, cross the 5mm sieve, every basin is adorned native 200g; The plantation peanut, 60%, 30 day post-sampling of adjusting water cut to maxmun field capacity; (LA1600+scanner is after Canada) scanning obtains the root system image, with root system analysis software (Winrhizo2003b with the root system scanner; Canada) carry out the related root index analysis, measure soil IAA content, and measure the soil mineral nitrogen with the HPLC method; Quick-acting potassium content, plant fresh weight, plant height and the complete full potassium content of nitrogen.
Peanut seed: peanut seed carries out 20% ydrogen peroxide 50 surface sterilization 20min, aseptic water washing repeatedly, vernalization 2d, it is subsequent use to choose the consistent seed that germinates.
Connecing bacterium handles: JX1 of the present invention (CGMCC No.5624) is inoculated in the LB liquid nutrient medium, and 30 ℃, 180rmin
-1Shaking table is cultivated, and culture bacteria is grown to logarithmic phase, then with bacteria suspension 10000rmin
-1Centrifugal 10min uses sterilized water resuspended again, operates triplicate equally, and bacteria suspension is evenly sprayed in soil, and inoculum size is 10
8CFUg
-1(be every gram dry ground inoculation 10
8CFU JX1).
Control treatment: as contrast, soil does not spray JX1 bacterium liquid, adds the equivalent sterilized water.
The result is shown in Fig. 7-17.Can find out inoculated the overground part fresh weight of the peanut plant that JX1 (CGMCC No.5624) soil grows, and plant height all has significantly rising tendency than CK by Fig. 7 and Fig. 8; Because JX1 (CGMCC No.5624) has the effect of fixed nitrogen potassium decomposing, make that mineral nitrogen and quick-acting potassium content increase in the soil (Figure 16, Figure 17); Thereby promoted plant to N, and the absorption of elements such as K (Fig. 9, Figure 10); Can find out that from Figure 11-14 inoculation JX1 handles and do not connect the bacterium processing and compares the peanut system general length; Root surface area, average root diameter and tip of a root number all significantly increase, and have promoted the growth of peanut root system; As can beappreciated from fig. 15, after connecing the bacterium processing, soil IAA content significantly increases, and exceeds about 2 times than control group.Can find out that in conjunction with above result the short living bacterium JX1 of rhizosphere of the present invention has positive effect to growth, the growth of foundation, IAA output is high, can effectively promote crop growth.
The above only is a preferred implementation of the present invention; Be noted that for those skilled in the art; Under the prerequisite that does not break away from the principle of the invention, can also make some improvement and retouching, these improvement and retouching also should be regarded as protection scope of the present invention.
Claims (4)
1. a rhizosphere is urged living bacterium JX1; Classification called after bacillus amyloliquefaciens (Bacillus amyloliquefaciens); On December 20th, 2011 was preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, and preserving number is CGMCC No.5624.
2. the described preserving number of claim 1 is the application of the short living bacterium JX1 of rhizosphere in promoting plant-growth of CGMCC No.5624.
3. application according to claim 2 is characterized in that described plant is a peanut.
4. the described preserving number of claim 1 is the short application of living bacterium JX1 in peanut cultivation of rhizosphere of CGMCC No.5624.
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| CN103952332A (en) * | 2014-03-14 | 2014-07-30 | 南京博农生物科技有限公司 | Bacillus amyloliquefaciens ZH1 and its application |
| CN104232538A (en) * | 2014-09-03 | 2014-12-24 | 南京聚肽高科农业有限公司 | Efficient potassium bacterium and application thereof |
| CN104232509A (en) * | 2014-07-25 | 2014-12-24 | 海南万绿宝生物科技有限公司 | Bacillus amyloliquefaciens bacterial strain Bg-1 and application thereof |
| CN104560789A (en) * | 2014-12-13 | 2015-04-29 | 郑州市污水净化有限公司 | Peanut growth promoting rhizobacteria HS2 and application thereof |
| CN104560785A (en) * | 2014-12-13 | 2015-04-29 | 郑州市污水净化有限公司 | Corn growth-promoting rhizobacteria YM11 and application thereof |
| CN104962492A (en) * | 2015-06-18 | 2015-10-07 | 中国农业大学 | Bacillus amyloliquefaciens, method for preparing solid inoculant thereof and application of solid inoculant |
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| CN109673469A (en) * | 2018-12-28 | 2019-04-26 | 云南云叶化肥股份有限公司 | A kind of preparation method with growth promotion seedling medium |
| CN110447494A (en) * | 2019-09-02 | 2019-11-15 | 广东海粤盛农业有限公司 | A kind of high yield cultivation method of peanut |
| CN111592992A (en) * | 2019-02-21 | 2020-08-28 | 中国科学院南京土壤研究所 | Preparation method and application of bacillus amyloliquefaciens microbial inoculum |
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| CN103952332A (en) * | 2014-03-14 | 2014-07-30 | 南京博农生物科技有限公司 | Bacillus amyloliquefaciens ZH1 and its application |
| CN103952332B (en) * | 2014-03-14 | 2016-08-24 | 南京博农生物科技有限公司 | A kind of bacillus amyloliquefaciens ZH1 and application thereof |
| CN104232509B (en) * | 2014-07-25 | 2016-09-21 | 海南万绿宝生物科技有限公司 | A kind of Bacillus amyloliquefaciens strain Bg-1 and application thereof |
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| CN105815338A (en) * | 2016-03-24 | 2016-08-03 | 吉林农业大学 | Biopesticide for ginseng seed and soil treatment and preparation method thereof |
| CN106011005A (en) * | 2016-05-20 | 2016-10-12 | 东莞市保得生物工程有限公司 | Bacillus amyloliquefaciens T600 and preparation method and application of microbial agent |
| CN106011005B (en) * | 2016-05-20 | 2020-01-17 | 东莞市保得生物工程有限公司 | Bacillus amyloliquefaciens T600 and preparation method and application of microbial inoculum thereof |
| CN108017447A (en) * | 2017-12-11 | 2018-05-11 | 河南邑鸿善成生物技术有限公司 | Prepare the method for bacillus amyloliquefaciens fermentation fertilizer and its application of fertilizer |
| CN109673469A (en) * | 2018-12-28 | 2019-04-26 | 云南云叶化肥股份有限公司 | A kind of preparation method with growth promotion seedling medium |
| CN111592992A (en) * | 2019-02-21 | 2020-08-28 | 中国科学院南京土壤研究所 | Preparation method and application of bacillus amyloliquefaciens microbial inoculum |
| CN110447494A (en) * | 2019-09-02 | 2019-11-15 | 广东海粤盛农业有限公司 | A kind of high yield cultivation method of peanut |
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